Buchbesprechungen

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Buchbesprechungen

Ohne Zusammenfassung     

A general method for highly selective cross-linking of unprotected polypeptides via pH-controlled modification of N-terminal alpha-amino groups

A method is described for the highly selective modification of the alpha-amino groups at the N-termini of unprotected peptides to form stable, modified peptide intermediates which can be covalently coupled to other molecules or to a solid support. Acylation with iodoacetic anhydride at pH 6.0 occurs with 90-98% selectivity for the alpha-amino group, depending on the N-terminal residue (as shown with a series of model hexapeptides containing a competing Lys residue). Although Cys residues must be protected (reversibly or irreversibly) before the anhydride reaction, there are no detectable side reactions of the alpha-amino moiety--of the reagent or of modified peptide--with the side chains of His, Met, or Lys. The reaction works well in denaturants, so that inhibitory effects of noncovalent structure can be minimized. In a second step the iodoacetyl-peptide can be reacted with a thiol group on a protein, on a solid chromatography matrix, on a spectroscopic probe, etc. This is illustrated by reaction of a series of N alpha-iodoacetyl-peptides with murine interferon-gamma, which contains a C-terminal Cys residue. Data are presented which suggest that this iodoacetic anhydride scheme is superior in selectivity for alpha-amino groups to conventional chemical approaches to cross-linking such as use of 2-iminothiolane or N-hydroxysuccinimide-activated carboxylic acid esters. The reaction is ideally suited for modifying peptide fragments, as pure species or as mixtures, derived from proteolytic or chemical fragmentation of proteins. Furthermore, polypeptides synthesized biosynthetically, for example via recombinant DNA techniques, can be cross-linked in this way.(ABSTRACT TRUNCATED AT 250 WORDS)     

Isolation and characterization of acetonitrile utilizing bacteria

Summary Bacteria utilizing high concentrations of acetonitrile as the sole carbon source were isolated and identified asChromobacterium sp. andPseudomonas aeruginosa. Maximum growth was attained after 96 h of incubation andP. aeruginosa grew slightly faster thanChromobacterium sp. The strains were able to grow and oxidize acetonitrile at concentrations as high as 600 mM. However, higher concentrations inhibited growth and oxygen uptake. Degradation studies with (¹⁴C)acetonitrile indicated 57% of acetonitrile was degraded byPseudomonas aeruginosa as compared to 43% byChromobacterium. The isolates utilized different nitrile compounds as carbon substrates.     

The xylem sap of maple (Acer platanoides) trees--sap obtained by a novel method shows changes with season and height

Xylem sap of log pieces of maple trees was collected by a novelpressure/decompression method developed recently for the mechanicaldrying of timber. Seasonal changes and spatial distributionsof the osmotic potential, the pH and the concentrations of majorsolutes and of the plant stress-hormone abscisic acid (ABA)were investigated. Sucrose and quebrachitol were the main components of the xylemsap. The sucrose concentration varied between 10 mM and 25 mMduring the winter months and declined to a minimum during theperiod of bud burst. Quebrachitol was found in concentrationsof up to 7 mM with a high variability throughout the year. Highconcentrations of ABA were measured during the summer seasonand in mid-winter. Rainfall caused an increase of ABA in somesamples. The osmotic potential of the xylem sap increased with the heightof the collection site. The pH of the sap decreased by approximatelyone unit between the base of the trunk and the crown. The increaseof the osmotic potential was mainly due to sucrose, quebrachitoland potassium. Malate contributed to the decrease of the pH.ABA of the xylem sap increased with decreasing moisture contentof the wood. Key words: Pressure/decompression method, xylem sap, abscisic acid, sucrose, quebrachitol, Acer platanoides     

Transformation of the extremely thermoacidophilic archaeon Sulfolobus solfataricus via a self-spreading vector

Abstract The comparative chromosomal locations of polymeric β-fructosidase SUC genes have been determined by Southern blot hybridization with the SUC2 probe in 91 different strains of Saccharomyces cerevisiae . Most of the strains exhibited a single SUC2 gene, but in some strains two or three SUC genes were found. All Suc⁻ strains carried a silent suc2⁰ sequence. The accumulation of SUC genes was observed in populations derived from sources containing sucrose and seems to be absent in strains from sources promoting the MEL gene.     

Cell Envelope Changes in Solvent-Tolerant and Solvent-Sensitive Pseudomonas putida Strains following Exposure to o-Xylene

Solvent-tolerant and -sensitive Pseudomonas putida strains were studied to determine their cell envelope changes following exposure to o-xylene. Both strains produced trans-unsaturated fatty acids. The tolerant strain showed an increase in total fatty acids, an increase in saturated fatty acids, and modified lipopolysaccharide. It is suggested that these envelope modifications aid in survival at high concentrations of organic solvents.     

The influence of the cytosolic oncotic pressure on the permeability of the mitochondrial outer membrane for ADP: implications for the kinetic properties of mitochondrial creatine kinase and for ADP channelling into the intermembrane space

Summary Cytosolic proteins as components of the physiological mitochondrial environment were substituted by dextrans added to media normally used for incubation of isolated mitochondria. Under these conditions the volume of the intermembrane space decreases and the contact sites between the both mitochondrial membranes increase drastically. These morphological changes are accompanied by a reduced permeability of the mitochondrial outer compartment for adenine nucleotides as it was shown by extensive kinetic studies of mitochondrial enzymes (oxidative phosphorylation, mi-creatine kinase, mi-adenylate kinase). The decreased permeability of the mitochondrial outer membrane causes increased rate dependent concentration gradients in the micromolar range for adenine nucleotides between the intermembrane space and the extramitochondrial space. Although all metabolites crossing the outer membrane exhibit the same concentration gradients, considerable compartmentations are detectable for ADP only due to its low extramitochondrial concentration. The consequences of ADP-compartmentation in the mitochondrial intermembrane space for ADP-channelling into the mitochondria are discussed.     

Nutrient additions by waterfowl to lakes and reservoirs: predicting their effects on productivity and water quality

Lakes and reservoirs provide water for human needs and habitat for aquatic birds. Managers of such waters may ask whether nutrients added by waterfowl degrade water quality. For lakes and reservoirs where primary productivity is limited by phosphorus (P), we developed a procedure that integrates annual P loads from waterfowl and other external sources, applies a nutrient load-response model, and determines whether waterfowl that used the lake or reservoir degraded water quality. Annual P loading by waterfowl can be derived from a figure in this report, using the days per year that each kind spent on any lake or reservoir. In our example, over 6500 Canada geese (Branta canadensis) and 4200 ducks (mostly mallards, Anas platyrhynchos) added 4462 kg of carbon (C), 280 kg of nitrogen (N), and 88 kg of P y^–1 to Wintergreen Lake in southwestern Michigan, mostly during their migration. These amounts were 69% of all C, 27% of all N, and 70% of all P that entered the lake from external sources. Loads from all external sources totaled 840 mg P m^–2 y^–1. Application of a nutrient load-response model to this concentration, the hydraulic load (0.25 m y^–1), and the water residence time (9.7 y) of Wintergreen Lake yielded an average annual concentration of total P in the lake of 818 mg m^–3 that classified the lake as hypertrophic. This trophic classification agreed with independent measures of primary productivity, chlorophyll-a, total P, total N, and Secchi disk transparency made in Wintergreen Lake. Our procedure showed that waterfowl caused low water quality in Wintergreen Lake.Contribution 824 of the National Fisheries Research Center-Great Lakes, 1451 Green Road, Ann Arbor, Michigan 48105, U.S.A. and 722 of the Kellogg Biological Station, Michigan State University.